Assay Detail
Binding
CHEMBL5734065
Review assay metadata, readout intent, target linkage, and publication context from the same page.
In Vitro JAK Kinase Assay: Compounds herein were tested for inhibitory activity of JAK targets according to the following in vitro assay described in Park et al., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (a.a. 837-1142), Jak2 (a.a. 828-1132) and Jak3 (a.a. 781-1124) with an N-terminal His tag were expressed using baculovirus in insect cells and purified. The catalytic activity of JAK1, JAK2 or JAK3 was assayed by measuring the phosphorylation of a biotinylated peptide. The phosphorylated peptide was detected by homogenous time resolved fluorescence (HTRF). IC50s of compounds were measured for each kinase in the 40 microL reactions that contain the enzyme, ATP and 500 nM peptide in 50 mM Tris (pH 7.8) buffer with 100 mM NaCl, 5 mM DTT, and 0.1 mg/mL (0.01%) BSA. For the 1 mM IC50 measurements, ATP concentration in the reactions was 1 mM. Reactions were carried out at room temperature for 1 hr and then stopped with 20 μL 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 in assay buffer (Perkin Elmer, Boston, Mass.). Binding to the Europium labeled antibody took place for 40 minutes and HTRF signal was measured on a Fusion plate reader (Perkin Elmer, Boston, Mass.).
1191
Total Activities
387
Compounds Tested
3
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Organism | Homo sapiens |
| Confidence | 9 — Direct single protein target |
| Curated By | Autocuration |
Publication
Piperidin-4-yl azetidine derivatives as JAK1 inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 397 | 7.46 | 7.60 |
| kon | 397 | - | - |
| k_off | 397 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL3977214 | — | — | 3 | 7.60 |
| CHEMBL3895594 | — | — | 3 | 7.52 |
| CHEMBL3915198 | — | — | 3 | 7.52 |
| CHEMBL3912772 | — | — | 3 | 7.52 |
| CHEMBL3961956 | — | — | 3 | 7.52 |
| CHEMBL5952192 | — | — | 3 | 7.52 |
| CHEMBL3924462 | — | — | 3 | 7.52 |
| CHEMBL3927154 | — | — | 3 | 7.52 |
| CHEMBL3914490 | — | — | 3 | 7.52 |
| CHEMBL3907279 | — | — | 3 | 7.52 |
| CHEMBL3943852 | — | — | 3 | 7.52 |
| CHEMBL3959500 | — | — | 3 | 7.52 |
| CHEMBL3980119 | — | — | 3 | 7.52 |
| CHEMBL3925310 | — | — | 3 | 7.52 |
| CHEMBL3931636 | — | — | 3 | 7.52 |
| CHEMBL3936342 | — | — | 3 | 7.52 |
| CHEMBL3982495 | — | — | 3 | 7.52 |
| CHEMBL3958002 | — | — | 3 | 7.52 |
| CHEMBL3934393 | — | — | 3 | 7.52 |
| CHEMBL3945897 | — | — | 6 | 7.52 |
| CHEMBL3936441 | — | — | 3 | 7.52 |
| CHEMBL3984476 | — | — | 3 | 7.52 |
| CHEMBL3929735 | — | — | 3 | 7.52 |
| CHEMBL3943023 | — | — | 3 | 7.52 |
| CHEMBL3986743 | — | — | 3 | 7.52 |
| CHEMBL3945039 | — | — | 3 | 7.52 |
| CHEMBL3973560 | — | — | 3 | 7.52 |
| CHEMBL3913894 | — | — | 3 | 7.52 |
| CHEMBL3928999 | — | — | 3 | 7.52 |
| CHEMBL3928468 | — | — | 6 | 7.52 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL3977214 | — | IC50 | = | 25.0 | nM | 7.60 |
| CHEMBL3903620 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3944041 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3936342 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3925174 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3931636 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL5910252 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3936845 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3926544 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3925310 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3897382 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3980119 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3954206 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3940789 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3912101 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3918254 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3976504 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3970498 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3973035 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3973120 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3923848 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3930325 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3933570 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3950173 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3952888 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL5893028 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3925297 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3892282 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3934393 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3971101 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3924438 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3889651 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3984158 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3939415 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL5783704 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL5894344 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3970818 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3972573 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3895594 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3905971 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3907114 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3908780 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3958002 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3982495 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3984668 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3954565 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL5935513 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3934692 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3895426 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL3974147 | — | IC50 | = | 30.0 | nM | 7.52 |